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J Chromatogr B Biomed Sci Appl. 1999 Sep 24;732(2):411-23.

Reversed-phase high-performance liquid chromatographic assay for the adenovirus type 5 proteome.

Author information

1
Berlex Laboratories, Inc., Richmond, CA 94804-0099, USA. elisabeth_lehmberg@Berlex.com

Abstract

An RP-HPLC assay was developed for a recombinant adenovirus type 5. During chromatography, intact adenovirus dissociated into its structural components (DNA and proteins) and the viral proteome was separated yielding a characteristic fingerprint. The individual components were identified by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy, N-terminal sequencing and amino acid composition. The assay was utilized to measure adenovirus particle concentration through quantification of structural proteins. Each structural protein provided independent measurement of virus concentration allowing verification of accuracy. The assay sensitivity is at or below 2 x 10(8) particles. Contrary to the benchmark spectrophotometric assay, the RP-HPLC assay was shown to be insensitive to contaminants common for partially purified adenovirus preparations.

PMID:
10517364
DOI:
10.1016/s0378-4347(99)00316-3
[Indexed for MEDLINE]

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